Fundeni Centre for Immunogenetics & Virology Telomerase and Hepsin as prognostic factors in prostate cancer progression and metastasis Ileana Constantinescu*, Rodica Tudor*** M.Harza**, M.Manu**, G.Gluck** and I. Sinescu** Centre for Immunogenetics and Virology* Centre of Urology, Dialysis, Nephrology and Kidney Transplantation** Fundeni Clinical Institute Batistei Medical Centre***

Fundeni Centre for Immunogenetics & Virology Pecorino L. Molecular Biology of Cancer, Mechanisms, Targets, and Therapeutics 2005, 1: 3, Oxford University Press.

The six hallmarks of cancer Fundeni Centre for Immunogenetics & Virology The six hallmarks of cancer Unlimited replicative potential Altered regulation of telomere maintenance results in unlimited replicative potential Angiogenesis (formation of new blood vessels) Invasion and metastasis The movement of cancer cells to other parts of the body is a major cause of cancer deaths Mutations alter the activity of enzymes involved in invasion and alter molecules involved in cell-cell and cetlextracellular adhesion Pecorino L. Molecular Biology of Cancer, Mechanisms, Targets, and Therapeutics 2005, 1: 3, Oxford University Press.

Fundeni Centre for Immunogenetics & Virology Introduction Prostate cancer is the most common male malignancy and the second leading cause of male cancer death; therefore is a great demand for noninvasive assay for early detection of prostate cancer. Working prostate tumor samples is very dificult because the malignancy is heterougenous and multifocal and early-stage tumors are nonpalpable. On the contrary, exfoliated cells representthe cancer status of the entire gland better due to the general tendency of cancer cells to exfoliate into biological fluids. One of the aim of this research is to clarify whether quantitative analysis of telomerase activity in exfoliated cells in urine could serve as a reliable molecular marker of prostate malignancy. The second aspect of interest is that prostate cancer related deaths are associated with metastasis. However, little is known about the mechanisms of this process. Hepsin is a cell surface serine protease that is markedly upregulated in human prostate cancer; however, the functional significance of this upregulationis unknown. There are scientific evidence that hepsin overexpression in prostate epithelium in vivo causes disorganization of the basement membrane. Botchkina G.I, Kim R.H, Botchkina I.L, Kirshenbaum A, Frisher Z, Adler H.L Noninvasive Detection of Prostate Cancer by Quantitative Analysis of Telomerase Activity, Clin Cancer Res, 2005;11 (9) 1.

Gene structure Fundeni Centre for Immunogenetics & Virology Pecorino L. Molecular Biology of Cancer, Mechanisms, Targets, and Therapeutics 2005, 1: 3, Oxford University Press.

Types of mutation Fundeni Centre for Immunogenetics & Virology Pecorino L. Molecular Biology of Cancer, Mechanisms, Targets, and Therapeutics 2005, 1: 3, Oxford University Press.

Fundeni Centre for Immunogenetics & Virology Material and Methods We have selected 58 patients with biopsy proven prostate cancer (N=38), with elevated total PSA levels and also with normal PSA levels. We have also included patients with PBH (N=20).There were included patients with (N=8) and without metastatsis (N=30). For the assessment of PSA we have used Microparticle Enzyme ImmunoAssay (MEIA) AxSym, ABBOTT. Telomerase was worked by PCR and Hepsin by real time PCR. Quantitative ELISA was used for the measurement of serum neopterin.

Fundeni Centre for Immunogenetics & Virology Results In all patients with elevated total PSA levels both telomerase and hepsin genes over expression were found. These values were directly correlated with significantly high serum of neopterin levels. In some patients with normal PSA levels we have noticed significantly raised telomerase gene expression. In some patients with normal PSA neopterin levels were normal. In patients with metastasis hepsin gene overexpression was noticed. A small percentage of patients without metastasis had hepsin overexpression as well.

HighTelomerase Prostate Cancer Activity (TA) 100% N=38 Fundeni Centre for Immunogenetics & Virology HighTelomerase Activity (TA) 100% Prostate Cancer N=38

Low TA 15% High TA 12% No Telomerase Activity 73 % Fundeni Centre for Immunogenetics & Virology Low TA 15% High TA 12% No Telomerase Activity 73 %

Fundeni Centre for Immunogenetics & Virology

Fundeni Centre for Immunogenetics & Virology

Fundeni Centre for Immunogenetics & Virology Conclusions Telomerase and hepsin are useful noninvasive markers for complete assessment of prostate cancer. Telomerase gene expression is useful for both early detection and diagnostic of prostate cancer. Hepsin is valuable for early detection of metastatic invasion in prostate cancer. Also, both markers are very useful as monitoring tools of anti cancer therapy in these patients.